Human breast tissues were fixed in 10% formalin overnight and then embedded in paraffin. Next, the tissue sections were dewaxed, rehydrated, and treated in citrate buffer and 3% hydrogen peroxide, respectively. And then incubated overnight at 4°C with anti- CES2 (antibody DF6433, 1:100, Affinity). The bound antibodies were detected using Secondary antibody binding to horseradish peroxidase and developed with 3,3’-diaminobenzidine using a two-step test kit (PV-9000, Zhongshan Jinqiao Biological).
Df6433
Manufactured by Affinity Biosciences
Sourced in United Kingdom
The DF6433 is a multi-channel digital pipette developed by Affinity Biosciences. It offers precise and accurate liquid handling for a wide range of volumes, enabling efficient and reliable sample management during various laboratory workflows.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using df6433
1
Immunohistochemical Analysis of CES2 in Breast Tissues
2
Quantitative Analysis of Hepatic and Intestinal Transporters
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The livers and jejunum tissue were homogenized in ice-cold radio immunoprecipitation assay (RIPA) buffer [50 mM Tris HCl, 1 mM EDTA, 150 mM NaCl, 0.1% sodium dodecyl sulfate, 0.5% sodium deoxycholate, 1.0% Nonidet P-40, and phenylmethane sulfonyl fluoride (PMSF), pH 8.0]. The homogenate was centrifuged at 12,000 rpm for 20 min at 4°C. The supernatants were collected for Western blot analysis, which was performed as described previously (Kong et al., 2015 (link)). The antibodies used in the present study included anti-Mrp-2 (1:500; sc-5770, Santa Cruz; California, USA), anti-glyceraldehyde 3-phosphate dehydrogenase (anti-GAPDH) (1:20,000; YM3029, Immunoway; Texas, USA), anti-UGT1A1 (1:4,000; ab194697, Abcam; Cambs, UK), anti-P glycoprotein (1:1,000; ab170904, Abcam; Cambs, UK), anti-CES2 (1:1,000; DF6433, Affinity; Cambs, UK), and anti-Bcrp (1:200; sc-69988, Santa Cruz; California, USA). The gray intensities of the bands were quantified using ImageJ software (National Institute of Health, MD, USA).
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